Resumo

Ganoderma lucidum is a medicinal mushroom widely recognized as a source of biomolecules with pharmacological properties, however, little is known about the factors that influence the synthesis of bioactive proteins by this fungus when cultivated under submerged fermentation. The objective of this work was to evaluate the production of mycelial biomass and intracellular proteases and protease inhibitors by G. lucidum cultivated under different submerged fermentation conditions. The cultivation was carried out in a medium composed of glucose (10 or 20 g.L-1), soy peptone (2.5 or 5 g.L-1) and yeast extract (5 g.L-1), with incubation under agitation (120 rpm) and non-agitation, totaling 8 experimental conditions. Biomass production was determined from the dry weight, while glucose consumption was estimated by quantification of reducing sugars. The proteins were extracted in NaCl (0.15 M), and the protein extracts were submitted to protein quantification by the Bradford method, total proteolytic activity using azocasein, caseinolytic and fibrinolytic activity in Petri dishes, activity of serine (trypsin and chymotrypsin) and cysteine (papain) protease inhibitors. Cultivation in agitated condition showed higher biomass production with a maximum value of 7 g.L-1, in addition to higher activities of trypsin, chymotrypsin and papain inhibitors, with 154 IU.mg-1, 153 IU.mg-1 e 343 IU.mg-1 of protein, respectively. The non-agitated condition showed a greater potential for obtaining proteins, total proteases, caseinolytic and fibrinolytic enzymes, with maximum values of 433 mg.g-1 of extract, 71 U.mL-1 of extract, 63.62 mm2 and 50.27 mm2, respectively. Thus, a medium composed of soy peptone, yest extract and glucose in a 1:2:4 proportion is recommended, under agitation to produce protease inhibitors, and the non-agitated condition when the target is, mainly caseinolytic and fibrinolytic enzymes.

Ganoderma lucidum é um cogumelo medicinal amplamente reconhecido como fonte de biomoléculas com propriedades farmacológicas, entretanto, pouco se conhece acerca dos fatores que influenciam a síntese de proteínas bioativas por esse fungo, quando cultivado sob fermentação submersa. O objetivo deste trabalho foi avaliar a produção de biomassa micelial e de proteases e inibidores de proteases intracelulares por G. lucidum cultivado em diferentes condições de fermentação submersa. O cultivo foi realizado em meio contendo glicose (10 ou 20 g.L-1), peptona de soja (2,5 ou 5 g.L-1) e extrato de levedura (5 g.L-1), com incubação sob agitação (120 rpm) ou não-agitado, totalizando 8 condições experimentais. A produção de biomassa foi determinada a partir do peso seco e o consumo de glicose a partir da quantificação de açúcares redutores. As proteínas foram extraídas em NaCl (0,15 M) e os extratos proteicos foram submetidos à quantificação de proteínas pelo método de Bradford, atividade proteolítica total usando azocaseína, atividade caseinolítica e fibrinolítica em placa de Petri, atividade de inibidores de serino-proteases (tripsina e quimotripsina) e cisteíno-protease (papaína). O cultivo em condição agitada apresentou maior produção de biomassa com valor máximo de 7g.L-1, além de maiores atividades de inibidores de tripsina, quimotripsina e papaína, com 154 UI.mg-1, 153 UI.mg-1 e 343 UI.mg-1 de proteína, respectivamente. A condição não-agitada demonstrou maior potencial para a obtenção de proteínas, proteases totais, enzimas caseinolíticas e fibrinolíticas, com valores máximos de 433 mg.g-1 de extrato, 71 U.mL-1 de extrato, 63,62 mm2 e 50,27 mm2, respectivamente. Assim, recomenda-se o meio composto de peptona de soja, extrato de levedura e glicose na proporção 1:2:4, em condição agitada para a produção de inibidores de proteases, e a condição não-agitada para a síntese de proteases, principalmente enzimas caseinolíticas e fibrinolíticas.

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The aim of this work was to evaluate the growth and the proximate compositionof the mycelium-based bocaiuva pulp with the edible mushroom Pleurotusostreatuson green bocaiuva flour added with different sources of nitrogen (urea, ammonium nitrate and sulfate ammonia). Growth was monitored by kinectics. At the end, the proximate composition of the best three treatments (dehydrated green bocaiuva pulp and water, T1; dehydrated green bocaiuva pulp and ammonium nitrate, T3; and green bocaiuva pulp/wheat bran and ammonium nitrate, T7) was determined. Ammonium nitrate was the nitrogen source that showed the greatest growth in both substrates (T3:8.33 cm and T7:7.67 cm) in relation to the other treatments (4.67 to 7.17 cm), with emphasis on the green bocaiuva pulp. The substrate with green bocaiuva pulp and water was the one that showed the highest growth (7.50 cm), which was close to the treatment with mixed substrate and ammonium nitrate (7.67 cm). The treatment with the green bocaiuva pulp and ammonium nitrate (T3) was highlighted due to its significant increase in proteins (9.42 g 100 g-1) and fibers (5.21 g 100 g-1), and decrease in carbohydrates (9.52 g 100 g-1), in comparison to the other treatments T7 (8.94, 2.16, and 5.99 g 100 g-1, respectively) and T1 (2.78, 4.33, and 2.28 g 100 g-1, respectively). The product obtained from the growth of P. ostreatusin green bocaiuva pulp presents promising perspectives to be utilized as raw material for the development of new food products with added nutritional value.

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Pleurotus albidus, a naturally growing species in the Amazon region, has been considered a promising source of milk-clotting proteases. The production of such enzymes using lignocellulosic residues is a sustainable alternative to replace mammalian rennet. The application of P. albidus milk-clotting proteases in cheese making has not yet been reported in the scientific literature. The aim of this study was to characterize the milk-clotting proteases of P. albidus and use these enzymes in the production of Minas frescal cheese. For the production of coagulating proteases, the mushroom was grown in açaí seeds supplemented with rice bran (10%, w/w). The parameters affecting the production of coagulant, such as inoculum size, fermentation time, initial pH of cultivation medium and age of the inoculum were evaluated. The coagulant extract obtained under optimal production conditions was evaluated for optimal pH and temperature, pH and temperature stability, effect of ions and inhibitors. Significant production of coagulating proteases was obtained under the following conditions: inoculum size (2.5%), fermentation time (10 days), initial pH of the cultivation medium (6), and inoculum age (10 days). The coagulant exhibited significant catalytic activity in pH 5.0 at 55°C, with stability at 45°C and was completely inhibited by iodoacetic acid. The milk-clotting proteases of P. albidus were efficient for making Minas frescal cheese that presented 55.0% of moisture, 20.0% of lipids and 17.20% of protein. Pleurotus albidus is a potential source of milk-clotting proteases that can be applied in dairy industry for production of fresh Minas frescal cheese.

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Edible mushrooms have a number of medicinal properties and this study aimed to investigate the antimicrobial activity of Pleurotus eryngiiDPUA1816 in metabolic broths after being grown in submerged cultivation. Mycelial fragments of pure P. eryngiiculture was inoculated in sweet potato culture medium and incubated at 150 rpm for 15 days at 25°C. Pleurotuseryngiiwas also cultivated for 18 days under the same conditions, the mycelial biomass was separated by filtration for quantification. The supernatant was used in the diffusion test in agar and performed against Escherichia coliCCCD-E005, Staphylococcus aureus CCCD-S009, Pseudomonas aeruginosaCCCD-P004, Candida albicansCCCD-CC001, Candida parapsilosis CCCD-CC004 and Candida tropicalisCCCD-CC002. The samples showed no inhibitory activity against bacteria, however they showed some activity againstC. albicans(12.17 mm), C. parapsilosis(27.67 mm) and C. tropicalis(13.67 mm). After being cultivated for 18 days, P. eryngiiwas able to inhibit all yeasts after 12 days of culture, with an inhibition halo of 29.33 mm at 16 days against C. parapsilosis. This study demonstrates the antifungal potential filtered liquids from P.eryngiicultivated in purple-skinned sweet potato culture medium, which suggests the possibility of the use of this species by the pharmaceutical industry as a natural source of biological action.

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Abstract The aim of this research was to evaluate the efficiency of aqueous alkali-treated Brachiaria straw for the cultivation of appropriate species of oyster mushroom. The substrate used in the cultivation of various Pleurotus spp. was soaked for 20 min by using two different procedures: (i) 0.52.0% Ca(OH)2 in 100 L water, and (ii) 50250 L water. As a result, 1% Ca(OH)2 dissolved in 100 L water and 3.5 kg of Brachiaria straw presented the best production. The most suitable species for the application of the present method were P. pulmonarius and P. sapidus. The success of this technique is directly related to the concentration of Ca(OH)2 and water, the species, and the origin and quality of raw material used as the substrate in the production of oyster mushroom.

Resumo

Abstract The aim of this research was to evaluate the efficiency of aqueous alkali-treated Brachiaria straw for the cultivation of appropriate species of oyster mushroom. The substrate used in the cultivation of various Pleurotus spp. was soaked for 20 min by using two different procedures: (i) 0.5-2.0% Ca(OH)2 in 100 L water, and (ii) 50-250 L water. As a result, 1% Ca(OH)2 dissolved in 100 L water and 3.5 kg of Brachiaria straw presented the best production. The most suitable species for the application of the present method were P. pulmonarius and P. sapidus. The success of this technique is directly related to the concentration of Ca(OH)2 and water, the species, and the origin and quality of raw material used as the substrate in the production of oyster mushroom.

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Pleurotus ostreatus is able to bioaccumulate several metals in its cell structures; however, there are no reports on its capacity to bioaccumulate iron. The objective of this study was to evaluate cultivation variables to increase iron bioaccumulation in P. ostreatus mycelium. A full factorial design and a central composite design were utilized to evaluate the effect of the following variables: nitrogen and carbon sources, pH and iron concentration in the solid culture medium to produce iron bioaccumulated in mycelial biomass. The maximum production of P. ostreatus mycelial biomass was obtained with yeast extract at 2.96 g of nitrogen L−1 and glucose at 28.45 g L−1. The most important variable to bioaccumulation was the iron concentration in the cultivation medium. Iron concentration at 175 mg L−1 or higher in the culture medium strongly inhibits the mycelial growth. The highest iron concentration in the mycelium was 3500 mg kg−1 produced with iron addition of 300 mg L−1. The highest iron bioaccumulation in the mycelium was obtained in culture medium with 150 mg L−1 of iron. Iron bioaccumulation in P. ostreatus mycelium is a potential alternative to produce non-animal food sources of iron.

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Abstract Lignocellulose is the most abundant environmental component and a renewable organic resource in soil. There are some filamentous fungi which developed the ability to break down and use cellulose, hemicellulose and lignin as an energy source. The objective of this research was to analyze the effect of three nitrogen resources (ammonium sulfate, saltpetre, soybean) in the holocellulolitic activity of Lentinula edodes EF 50 using as substrate sawdust E. benthamii. An experimental design mixture was applied with repetition in the central point consisting of seven treatments (T) of equal concentrations of nitrogen in ammonium sulfate, potassium nitrate and soybean. The enzymatic activity of avicelase, carboxymetilcellulase, β-glucosidase, xylanases and manganese peroxidase was determined. The humidity, pH, water activity (aw) and qualitative analysis of mycelial growth in 8 times of cultivation were evaluated. The results showed negative effect on enzyme production in treatments with maximum concentration of ammonium sulfate and potassium nitrate. The treatments with cooked soybean flour expressed higher enzymatic activities in times of 3, 6 and 9 days of culture, except in the activity of manganese peroxidase. The highest production was observed in the treatment with ammonium sulfate, and soybean (83.86 UI.L–1) at 20 days of cultivation.

Resumo Lignocelulose é o componente mais abundante do meio ambiente e recurso orgânico renovável no solo. Alguns fungos filamentosos têm desenvolvido a habilidade de degradar e utilizar celulose, hemicelulose e lignina como fonte de energia. O objetivo deste trabalho foi analisar o efeito de três fontes de nitrogênio (sulfato de amônio, nitrato de potássio e farelo de soja) na atividade enzimática de Lentinula edodes EF 50 utilizando como substrato serragem de E. benthamii. Foi aplicado um planejamento experimental de mistura com três repetições no ponto central constituído de sete tratamentos (T) de iguais concentrações em nitrogênio de sulfato de amônia, nitrato de potássio e farinha de soja cozida. Foram determinadas a atividade enzimática da avicelase, carboximetilcelulase, β-glicosidase, xilanases e manganês peroxidase. Foram avaliados o teor de umidade, pH, atividade de água (aw) e análise qualitativa do crescimento micelial em 8 tempos de cultivo. Os resultados mostraram efeito negativo na produção das enzimas nos tratamentos com máxima concentração de sulfato de amônia e nitrato de potássio. Os tratamentos com farinha de soja cozida expressaram maiores atividades enzimáticas, nos tempos de 3, 6 e 9 dias de cultivo exceto na atividade do manganês peroxidase. A maior produção foi observada no tratamento com sulfato de amônia e farinha de soja cozida (83.86 UI.L–1) em 20 dias de cultivo.

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O objetivo deste trabalho foi avaliar a composição mineral (macro e micronutrientes) dos substratos [(inicial e residual (pós-colheita)] à base de diferentes combinações de resíduos (folha, pseudocaule e pseudocaule + folha) e cultivares de bananeira - Musa spp. (Thap Maeo, Prata Anã, Pelipita e Caipira), durante 49 dias de cultivo da linhagem POS 09/100 de Pleurotus ostreatus. Verificaram-se que todos os substratos à base de resíduos de diferentes cultivares de bananeira apresentaram quantidades satisfatórias de nutrientes para o cultivo de P. ostreatus, tanto na fase inicial de cultivo como na final.(AU)

The objective of this study was to evaluate the mineral composition (macro e micronutrients) of the substrates [initial and residual (postharvest)] based on different combinations of waste (leaf, pseudo-stem and pseudo-stem + leaf) and banana cultivars - Musa spp. (Thap Maeo, Prata Anã, Pelipita and Caipira) during 49 days for the cultivation of POS 09/100 strain of P. ostreatus. It was verified that all of the substrates based on different combinations of waste and banana cultivars presented satisfactory amounts of nutrients for the cultivation of P. ostreatus, both in the initial phase of cultivation and in the end.(AU)

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O cogumelo Agaricus bisporus é um cogumelo de clima temperado que requer baixa temperatura para a indução da frutificação. Essa exigência limita o seu cultivo no Brasil nos meses mais frios do ano ou exige a utilização de ambiente controlado, onerando muito a produção. Por isso, este trabalho teve como objetivo avaliar a produtividade de diferentes linhagens em função da temperatura de cultivo após a indução da frutificação. Diferentes linhagens de A. bisporus foram avaliadas quanto ao efeito da temperatura sobre a colonização do composto e a produtividade do cogumelo. As temperaturas de 21 e 25°C foram utilizadas durante a colonização do composto e, após a indução da frutificação, durante o ciclo de cultivo. Para todas as linhagens testadas, a temperatura de 25°C proporcionou maior velocidade de colonização do composto, favorecendo ciclos de cultivo mais curtos. Além disso, após a indução da frutificação a 18°C, a manutenção do ambiente de cultivo a 25°C proporcionou maior produtividade, quando comparada ao cultivo a 21°C. Para a linhagem mais produtiva, os três primeiros fluxos foram responsáveis por mais de 50% da produção total. Portanto, para um cultivo comercial em que é necessário manter volumes constantes de produção, os ciclos de cultivo poderiam ser limitados a três fluxos de produção.

The Agaricus bisporus is a button mushroom that requires low temperature to induce fruiting. This requirement limits our cultivation in Brazil during the coldest months of the year and requires the use of controlled environment. Therefore, this study aimed to assess the productivity of different strains depending on the growth temperature after induction of fruiting. A. bisporus strains were evaluated for the effect of temperature on the colonization of compost and mushroom productivity. Temperatures of 21 and 25°C were used for the substrate colonization and after induction of fruiting crop cycle. For all strains tested, temperature 25°C gave a quickly colonization of the compound, favoring shorter production cycles. Furthermore, after induction of fruiting at 18°C, maintaining the culture environment at 25°C gave a higher productivity to contrast with culture at 21°C. For the most productive strain, the first three streams were responsible for over 50% of total production. Thus, for a cash crop where it is necessary to maintain constant volumes of production, crop cycles could be limited to three productions flows.

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Trichoderma spp is the cause of the green mold disease in mushroom cultivation production. Many disinfection treatments are commonly applied to lignocellulose substrates to prevent contamination. Mushroom growers are usually worried about the contaminations that may occur after these treatments during handling or spawning. The aim of this paper is to estimate the growth of the green mold Trichoderma sp on lignocellulose substrates after different disinfection treatments to know which of them is more effective to avoid contamination during spawning phase. Three different treatments were assayed: sterilization (121 ºC), immersion in hot water (60 and 80 ºC), and immersion in alkalinized water. Wheat straw, wheat seeds and Eucalyptus or Populus sawdust were used separately as substrates. After the disinfection treatments, bagged substrates were sprayed with 3 mL of suspension of conidia of Trichoderma sp (10(5) conidia/mL) and then separately spawned with Pleurotus ostreatus or Gymnopilus pampeanus. The growth of Trichoderma sp was evaluated based on a qualitative scale. Trichoderma sp could not grow on non-sterilized substrates. Immersions in hot water treatments and immersion in alkalinized water were also unfavorable treatments for its growth. Co- cultivation with mushrooms favored Trichoderma sp growth. Mushroom cultivation disinfection treatments of lignocellulose substrates influence on the growth of Trichoderma sp when contaminations occur during spawning phase. The immersion in hot water at 60 ºC for 30 min or in alkalinized water for 36 h, are treatments which better reduced the contaminations with Trichoderma sp during spawning phase for the cultivation of lignicolous species.

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The gender Pleurotus is also known as oyster mushroom, shimeji or hiratake. Aiming to select the best substrates to cultivate two species of Pleurotus, this work measured vigor, mycelium growth (cm.day-1), fresh mass (g), productivity (%) and biological efficiency (%) of P. sajor-caju (PSC96/03) and P. ostreatoroseus (POR01/06) cultivated in the following substrates: sugarcane bagasse, elephant grass, waste of castor oil plant and pasteurized rice straw. Fungal cultures were recovered in culture medium CDA. For the evaluation of mycelium growth, moist substrates were put into a closed assay tube with sterilized aluminum paper. Then, they were inoculated in 10 mm culture dishes and taken to the incubator at 26 ± 2C. Mycelium vigor was measured with grades from 1 to 3 according to density. For axenic cultivation, substrates were placed into 250 g flasks of substrate and autoclaved twice at 121C (1 atm) for 60 minutes, and then inoculated with 3% of spawn. The lineage P. sajor-caju (PSC96/03) showed higher growth rates in relation to P. ostreatoroseus (POR01/06). Substrates showing lower C/N ratio provided more mycelium vigor. Castor oil plant waste based-substrate showed good perspectives to growing P. sajor-caju (PSC96/03).

O gênero Pleurotus é conhecido como cogumelo ostra, shimeji ou hiratake. Neste trabalho, com a finalidade de selecionar substratos para o cultivo de duas espécies de Pleurotus, foram avaliados vigor, crescimento micelial (cm.dia-1), massa fresca (g), produtividade (%) e eficiência biológica (%) de P. sajor-caju (PSC96/03) e P. ostreatoroseus (POR01/06) cultivados nos substratos: bagaço de cana-de-açúcar, capim-elefante, resíduos da cultura da mamona e palha de arroz esterilizados. As culturas fúngicas foram recuperadas em meio de cultura CDA. Para avaliar o crescimento micelial, os substratos úmidos foram acondicionados em tubos de ensaio e fechados com papel alumínio esterilizados. Depois, foram inoculados com discos de cultura de 10 mm de diâmetro e incubados em estufa a 26 ± 2C. O vigor do micélio foi mensurado por meio de notas de 1 a 3, conforme adensamento. Para o cultivo axênico, os substratos foram colocados em frascos de 250 g de substrato e autoclavados duas vezes a 121C (1 atm) por 60 minutos, posteriormente inoculados com 3% de spawn. Ao serem analisadas, a linhagem P. sajor-caju (PSC96/03) apresentou maior velocidade de crescimento que a P. ostreatoroseus (POR01/06), os substratos com menor relação C/N propiciaram maior vigor de micélio e o substrato à base de resíduos da cultura da mamona apresentou perspectivas para o cultivo de P. sajor-caju (PSC96/03).

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Pleurotus species secrete phenol oxidase enzymes: laccase (Lcc) and manganese peroxidase (MnP). New genotypes of these species show potential to be used in processes aiming at the degradation of phenolic compounds, polycyclic aromatic hydrocarbons and dyes. Hence, a screening of some strains of Pleurotus towards Lcc and MnP production was performed in this work. Ten strains were grown through solid-state fermentation on a medium based on Pinus spp. sawdust, wheat bran and calcium carbonate. High Lcc and MnP activities were found with these strains. Highest Lcc activity, 741 ± 245 U gdm-1 of solid state-cultivation medium, was detected on strain IB11 after 32 days, while the highest MnP activity occurred with strains IB05, IB09, and IB11 (5,333 ± 357; 4,701 ± 652; 5,999 ± 1,078 U gdm-1, respectively). The results obtained here highlight the importance of further experiments with lignocellulolytic enzymes present in different strains of Pleurotus species. Such results also indicate the possibility of selecting more valuable strains for future biotechnological applications, in soil bioremediation and biological biomass pre-treatment in biofuels production, for instance, as well as obtaining value-added products from mushrooms, like phenol oxidase enzymes.

Resumo

The objective of this research was to evaluate the oyster mushroom Pleurotus ostreatus- (Jacq.: Fr.) Kumm. cultivation in substrates based on different combinations of wastes (leaf, pseudo-stem and pseudo-stem + leaf) and banana cultivars - Musa spp. (Thap Maeo, Prata Anã, Pelipita and Caipira) during 49 days. Organic matter loss in the substrate by action of the fungus was also evaluated during that period. It was verified that the pseudo-stem waste provided the best averages of biological efficiency among all cultivars tested and best rates were obtained by Thap Maeo (61.5%). The highest organic matter loss (OML) was obtained from pseudo-stem + leaf wastes (Prata Anã - 78.6%; Thap Maeo - 67.6%; Pelipita - 64.8%; Caipira - 60.6%). Therefore, the use of those wastes showed itself viable for P. ostreatus cultivation due to its availability and low cost, besides decreasing discards to environment.

Resumo

A determinação de biomassa micelial fúngica crescida em substratos de cultivo sólido particulado (SCSP) é ainda um desafio devido à dificuldade de separação do micélio e o substrato. O objetivo deste trabalho foi avaliar a técnica de microscopia de epifluorescência para determinação da biomassa micelial de Pleurotus ostreatus em SCSP. Para determinação da exatidão da metodologia P. ostreatus foi crescido em meio líquido de extrato de malte e; a biomassa micelial foi separada por centrifugação, liofilizada e moída. Concentrações conhecidas do pó do micélio foram misturadas ao SCSP, composto de bagaço de cana de açúcar e fibra de soja, previamente autoclavado. Em seguida, a biomassa micelial foi determinada por microscopia de epifluorescência. Para promover a variação da biomassa micelial a ser determinada por microscopia de epifluorescência, SCSP adicionado de diferentes concentrações de ferro foram utilizados para o crescimento do fungo. Concluiu- -se que a técnica apresenta baixa precisão e exatidão, o que implica na necessidade de maiores estudos para aplicação desta técnica para a determinação de biomassa micelial crescida em SCSP.

Determination of fungal mycelial biomass grown on solid substrate cultivation (SSC) is still a challenge due to the difficulty in separating mycelium and substrate. The objective of this study was to evaluate the epifluorescence microscopy to determine the mycelial biomass of Pleurotus ostreatus grown in SSC. P. ostreatus was grown in malt extract liquid medium and mycelial biomass was separated by centrifugation. It was then lyophilized and milled. Mycelial powder was added at known concentrations to SSC composed of sugarcane bagasse and soy fiber, previously autoclaved. Mycelial biomass was determined by epifluorescence microscopy. In order to promote mycelial biomass variation for the determination by epifluorescence microscopy, SSC added at different iron concentrations was used for fungus growth. It was concluded that the technique has low precision and accuracy, which implies the need for further studies in order to apply this technique for the determination of mycelial biomass grown in SSC.

La determinación de biomasa micelial fúngica crecida en sustratos de cultivo sólido particulado (SCSP) es todavía un reto debido a la dificultad de separación del micelio y el sustrato. El objetivo de este estudio fue evaluar la técnica de microscopía de epifluorescencia para determinación de la biomasa micelial de Pleurotus ostreatus en SCSP. Para determinación de exactitud de la metodología P. ostreatus se cultivó en medio líquido de extracto de malta y; la biomasa micelial separada por centrifugación, liofilizada y molida. Concentraciones conocidas del polvo del micelio fueron mezcladas al SCSP, compuesto de bagazo de caña de azúcar y fibra de soya, previamente tratado en autoclave. A continuación, la biomasa micelial a ser determinada por microscopía de epifluorescencia. Para promover la variación de la biomasa micelial a ser determinada por microscopía de epifluorescencia, SCSP añadida con diferentes concentraciones de hierro fueron utilizados para el crecimiento del hongo. Se concluye que la técnica presenta baja precisión y exactitud, lo que implica en la necesidad de realizar más estudios para aplicación de esta técnica para la determinación de biomasa micelial crecida en SCSP.

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Cascas de café são fonte de carboidratos e nutrientes que podem ser bioconvertidos em produtos de interesse como enzimas. Lacases são cobre polifenol oxidases que oxidam compostos fenólicos, enquanto reduzem oxigênio molecular à água e; sua baixa especificidade a substratos permite sua aplicação em várias áreas como indústria têxtil, de alimentos e biorremediação. Os objetivos desse trabalho foram avaliar a capacidade de produção de lacase de três linhagens de fungos basidiomicetos (Lentinula edodes U6/1, Pleurotus ostreatus U6/9 e Pleurotus florida U6/10) por fermentação submersa com cascas de café e avaliar o uso de cobre como indutor dessa enzima. A casca de café mostrou ser um bom substrato para produção de lacases e das três linhagens testadas Pleurotus ostreatus (U6/9) foi a mais produtiva (22,5 U mL-1). A melhor fonte de nitrogênio para produção de lacases de Pleurotus ostreatus (U6/9) foi o extrato de levedura na concentração de 9 g/L (20 U mL-1). A adição de 150 μM de CuSO4 resultou na indução significativa na produção de lacases nessa linhagem (21 U mL-1) no 12° dia de cultivo.

Coffee husks are a source of carbohydrates and nutrients that may be bioconverted into products of interest, such as enzymes. Laccases are copper polyphenol oxidases that oxidize phenolic compounds while reducing molecular oxygen to water. Laccase’s low specificity to substrates allows its application in several areas such as textiles, food processing and bioremediation industries. The aims of this study were to evaluate the potential to produce laccase from three strains of basidiomycetous fungi (Lentinula edodes U6/1, Pleurotus ostreatus U6/9, and Pleurotus florida U6/10) by submerged fermentation with coffee husks, and to evaluate the use of copper as an inducer of the enzyme. Coffee husk proved to be a good substrate for laccase production, with Pleurotus ostreatus (U6/9) being the most productive strain (22.5 U mL-1). The best source of nitrogen for laccase production of Pleurotus ostreatus (U6/9) was yeast extract 9 g/L (20 U mL-1). The addition of CuSO4 (150 μM) resulted in significant induction of laccase (21 U mL-1) on the 12th day of cultivation.

Cáscaras de café son fuente de carbohidratos y nutrientes que pueden ser bioconvertidos en productos de interés, tales como enzimas. Lacases son cobre polifenol oxidasas que oxidan compuestos fenólicos, mientras reducen el oxígeno molecular a el agua y; su baja especificidad a sustratos permite su aplicación en diversas áreas, como la industria textil, de alimentos y de biorremediación. Los objetivos de este estudio fueron evaluar la capacidad de producción de lacase de tres linajes de hongos basidiomicetos (Lentinula edodes U6/1, Pleurotus ostreatus U6/9 y Pleurotus florida U6/10) por fermentación sumergida con cáscaras de café, y evaluar el uso del cobre como inductor de esta enzima. La cáscara de café resultó ser un buen sustrato para la producción de lacases y, de las tres linajes probadas, Pleurotus ostreatus (U6/9) fue la más productiva (22,5 U mL-1). La mejor fuente de nitrógeno para la producción de lacases de Pleurotus ostreatus (U6/9) fue el extracto de levadura a una concentración de 9 g L-1 (20 U mL-1). La adición de 150 μM de CuSO4 resultó en la inducción significativa de Cáscaras de café son fuente de carbohidratos y nutrientes que pueden ser bioconvertidos en productos de interés, evadura a una concentración de 9 g L-1 (20 U mL-1). La adición de 150 μM de CuSO4 resultó en la inducción significativa de la producción de lacases en esa linaje (21 U mL-1), en el 12º día de cultivo.

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This study aimed to verify the biological efficiency and production flushes of Agaricus blazei strains on different casing layers during 90 cultivation days. Four casing layers were used: mixture of subsoil and charcoal (VCS), lime schist (LSC), São Paulo peat (SPP) and Santa Catarina peat (SCP); and two genetically distant A. blazei strains. The fungus was grown in composted substratum and, after total colonization, a pasteurized casing layer was added over the substratum, and fructification was induced. Mushrooms were picked up daily when the basidiocarp veil was stretched, but before the lamella were exposed. The biological efficiency (BE) was determined by the fresh basidiocarp mass divided by the substratum dry mass, expressed in percentage. The production flushes were also determined over time production. The BE and production flushes during 90 days were affected by the strains as well as by the casing layers. The ABL26 and LSC produced the best BE of 60.4 percent. Although VCS is the most used casing layer in Brazil, it is inferior to other casing layers, for all strains, throughout cultivation time. The strain, not the casing layer, is responsible for eventual variations of the average mushroom mass. In average, circa 50 percent of the mushroom production occurs around the first month, 30 percent in the second month, and 20 percent in third month. The casing layer water management depends on the casing layer type and the strain. Production flush responds better to water reposition, mainly with ABL26, and better porosity to LSC and SCP casing layers.

Resumo

Este trabalho objetivou avaliar a viabilidade do cultivo de Pleurotus sajor-caju utilizando diversos resíduos gerados na região de Pelotas, RS. Os substratos foram formulados com bagaço de cana-de-açúcar, capim-elefante e palha de arroz, utilizando-os separados e misturados entre si, totalizando sete tratamentos. Os substratos foram umedecidos por 24 horas, pasteurizados a 100º C durante 30 minutos, sendo em seguida adicionado de 3% de "spawn" de P. sajor-caju (PSC96/03) e acondicionado em embalagens de polietileno de 1 kg. A incubação foi a 25-28º C, durante 25 dias e a colheita ocorreu em dois fluxos durante 45 dias cada. Os cogumelos foram coletados e pesados para a avaliação da massa fresca (g), produtividade (%) e eficiência biológica (%). Os resultados mostraram que a palha de arroz produz mais no primeiro fluxo e o capim-elefante no segundo, porém, no somatório dos dois fluxos para massa fresca e produtividade, destaca-se o cultivo de P. sajor-caju no capim-elefante e a eficiência biológica na palha de arroz, sendo ambos, separadamente, substratos viáveis para o cultivo de cogumelos. O bagaço de cana-de-açúcar foi o substrato menos indicado para o cultivo deste cogumelo por esta técnica.

This research was aimed at evaluating the viability of the Pleurotus sajor-caju crop using various residues generated in the region of Pelotas, state of Rio Grand do Sul, Brazil. The substrates were formulated with sugarcane bagasse, elephant grass and rice straw, separate and mixed, for a total of 7 treatments. The substrates were moistened for 24 hours, pasteurized at 100º C for 30 minutes, and then seeded with a 3% spawn of P. sajor-caju (PSC96/03) and placed into 1 kg polyethylene bags. The incubation was carried out at 25-28º C, for 25 days, and the harvest took place in two flushes after periods of 45 days each. The mushrooms were collected and weighed for the evaluation of the fresh mass (g), productivity (%) and biological efficiency (%). The results showed that the rice straw produces more in the first flush and the elephant grass more in the second, while the overall totals for fresh mass and productivity of both flushes showed that each of these materials was, separately, a viable substrate for the cultivation of P. sajor-caju mushrooms. The sugarcane bagasse proved to be the least indicated substrate for the cultivation of this mushroom by means of this technique.

Resumo

O objetivo do trabalho foi avaliar o crescimento micelial, em placa de Petri, de dois fungos comestíveis (Pleurotus ostreatus e Lentinula edodes) em seis meios de cultura [(malte-ágar, serragemdextrose-ágar-marupá (SDA-MA), serragem-dextrose-ágar-cajuí (SDA-CA), serragem-dextroseágar-açaí (SDA-AÇA), serragem-dextrose-ágar-banana 50% (BAN 50%) e serragem-dextroseágar-banana 100% (BAN 100%)]. O delineamento experimental foi inteiramente casualizado, em esquema fatorial 6 x 2. Cada tratamento constou de seis repetições, correspondente a uma placa de Petri, totalizando 72 unidades experimentais. Verificou-se que, em todos os meios à base de resíduos, o P. ostreatus apresentou um melhor desenvolvimento micelial (81,00; 64,66; 81,00; 50,16; e 33,33 mm, para SDA-MA, SDA-CA, SDA-AÇA, BAN 50% e BAN 100%, respectivamente) que o L. edodes (32,00; 31,66; 27,66; 37,33; e 21,83 mm, para SDA-MA, SDA-CA, SDA-AÇA, BAN 50% e BAN 100%, respectivamente). Também constatou-se que, para os L. edodes, não houve vantagem, em relação ao crescimento micelial, no uso de meios à base de resíduos comparado ao meio malteágar (testemunha), o qual obteve o melhor desempenho (62,17mm). Já para o P. ostreatus, os meios SDA-MA e SDA-AÇA apresentaram as maiores médias de crescimento (81 mm), o que representa um incremento de crescimento de 34% em relação ao meio testemunha (malte-ágar), cujo média de crescimento foi de 60,33mm. Assim, de uma forma geral, os resíduos testados indicam potencial de aproveitamento na fungicultura, especialmente para o cultivo de P. ostreatus.

The objective of this work was to evaluate the mycelial growth of 2 edible fungi (Pleurotus ostreatus and Lentinula edodes) in 6 culture media [(malt-agar, sawdustdextrose-agar-marupá (SDA-MA), sawdust-dextrose-agar-cajuí (SDA-CA), sawdust-dextrose-agaraçaí (SDA-AÇA), sawdust-dextrose-agar-banana 50% (BAN 50%) and sawdust-dextrose-agar-banana 100% (BAN 100%)], in Petri dishes. The experimental design was totally randomized, in a 6x2 factorial scheme. Each treatment consisted of six repetitions in 1 Petri dish, totaling 72 experimental units. It was verified that P. ostreatus presented better mycelial development (81.00; 64.66; 81.00; 50.16 and 33.33mm for SDA-MA, SDA-CA, SDA-AÇA, BAN 50% and BAN 100%, respectively) than L. edodes (32.00; 31.66; 27.66; 37.33 and 21.83mm for SDA-MA, SDA-CA, SDA-AÇA, BAN 50% and BAN 100%, respectively). It was also verified that there was no advantage for L. edodes in relation to mycelial growth, when media based on residues were used, compared to malt-agar medium (control), which obtained the best performance (62.17mm). As for P. ostreatus, SDA-MA and SDA-AÇA medium presented the highest growth averages (81 mm), representing a growth increase of 34% in relation to the control medium (malt-agar), whose growth average was 60.33mm. Thus, the residues tested present potential to be used in fungiculture, especially for the cultivation of P. ostreatus.

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Fourteen strains of Grifola frondosa (Dicks.) S. F. Gray, originating from different regions (Asia, Europe and North America) were tested for lignin degradation, ligninolytic enzyme activities, protein accumulation and exopolysaccharide production during 55 days of cultivation on oak sawdust. Lignin degradation varied from 2.6 to7.1 percent of dry weight of the oak sawdust substrate among tested strains. The loss of dry matter in all screened fungi varied between 11.7 and 33.0 percent, and the amount of crude protein in the dry substrate varied between 0.94 to 2.55 percent. The strain, MBFBL 596, had the highest laccase activity (703.3 U/l), and the maximum peroxidase activity of 22.6 U/l was shown by the strain MBFBL 684. Several tested strains (MBFBL 21, 638 and 662) appeared to be good producers of exopolysaccharides (3.5, 3.5 and 3.2 mg/ml respectively).